Cell light eu apollo488 rna imaging kit

Cell-Light EU Apollo-488 RNA Imaging Kit (Ribobio, Catalog# C10316-3) was used. Cells were seeded in 15 mm glass-bottom confocal dishes at 10,000 cells per dish. Twenty-four hours later, cells were treated with chemicals and incubated with 0.5 mM ethyl uridine (EU) for 1 h. Cells were washed thoroughly and fixed after EU incubation. EU was detected using the imaging kit and the cells were stained with antibodies and other fixed-cells stains.

5-Ethynyluridine (EU) incorporation was performed by using Cell-Light EU Apollo488 RNA Imaging Kit (RiboBio). Briefly, cells were incubated in complete culture medium containing 500 μM EU for 1 hour before washing with PBS and fixed. The cells were stained with 0.5 μg/ml 4’,6-diamidino-2-phenylindole (DAPI) for 5 min and mounted in anti-fade solution. Image stacks were obtained by using the fluorescence microscopy (Olympus). Nucleoplasm regions were identified based on DNA (DAPI) staining. The median of each cell’s mean intensity of the extracted nuclear signals after background subtraction (the signals outside nuclei) were plotted and calculated by applying the Image J software.